In this section
@article{Climent-Catala:2023:10.1021/acssynbio.3c00599,
author = {Climent-Catala, A and Casas-Rodrigo, I and Iyer, S and Ledesma-Amaro, R and Ouldridge, TE},
doi = {10.1021/acssynbio.3c00599},
journal = {ACS Synthetic Biology},
pages = {3754--3765},
title = {Evaluating DFHBI-responsive RNA light-up aptamers as fluorescent reporters for gene expression},
url = {http://dx.doi.org/10.1021/acssynbio.3c00599},
volume = {12},
year = {2023}
}
TY - JOUR
AB - Protein-based fluorescent reporters have been widely used to characterize and localize biological processes in living cells. However, these reporters may have certain drawbacks for some applications, such as transcription-based studies or biological interactions with fast dynamics. In this context, RNA nanotechnology has emerged as a promising alternative, suggesting the use of functional RNA molecules as transcriptional fluorescent reporters. RNA-based aptamers can bind to nonfluorescent small molecules to activate their fluorescence. However, their performance as reporters of gene expression in living cells has not been fully characterized, unlike protein-based reporters. Here, we investigate the performance of three RNA light-up aptamersF30-2xdBroccoli, tRNA-Spinach, and Tornado Broccolias fluorescent reporters for gene expression in Escherichia coli and compare them to a protein reporter. We examine the activation range and effect on the cell growth of RNA light-up aptamers in time-course experiments and demonstrate that these aptamers are suitable transcriptional reporters over time. Using flow cytometry, we compare the variability at the single-cell level caused by the RNA fluorescent reporters and protein-based reporters. We found that the expression of RNA light-up aptamers produced higher variability in a population than that of their protein counterpart. Finally, we compare the dynamical behavior of these RNA light-up aptamers and protein-based reporters. We observed that RNA light-up aptamers might offer faster dynamics compared to a fluorescent protein in E. coli. The implementation of these transcriptional reporters may facilitate transcription-based studies, gain further insights into transcriptional processes, and expand the implementation of RNA-based circuits in bacterial cells.
AU - Climent-Catala,A
AU - Casas-Rodrigo,I
AU - Iyer,S
AU - Ledesma-Amaro,R
AU - Ouldridge,TE
DO - 10.1021/acssynbio.3c00599
EP - 3765
PY - 2023///
SN - 2161-5063
SP - 3754
TI - Evaluating DFHBI-responsive RNA light-up aptamers as fluorescent reporters for gene expression
T2 - ACS Synthetic Biology
UR - http://dx.doi.org/10.1021/acssynbio.3c00599
UR - https://www.ncbi.nlm.nih.gov/pubmed/37991880
UR - https://pubs.acs.org/doi/10.1021/acssynbio.3c00599
VL - 12
ER -
For any enquiries about the Fungal Science Network at Imperial, please contact: